ABSTRACT The DNA damage repair mechanism is highly dynamic, with many interacting components. However, it remains unclear how various repair modules dynamically interact. A greater understanding of these mechanisms could allow us to maximally leverage these dynamics for therapeutic benefit. I have developed an innovative high-throughput screening technique, Genome-Wide Dynamic Evaluation (GEoDE), that couples high-density 6,144 colony yeast colony genetic mutant arrays with time-lapse imaging to achieve genome- wide, time-resolved fitness screens which may be used to screen multiple therapeutic agents against thousands of colonies in real time. Analysis of the UV-induced DNA damage response using this technique has elucidated the dynamic sequencing of UV response pathways in yeast and identified 175 novel UV-responding genes. Importantly, GEoDE can identify the time at which each gene is needed in the recovery from UV- induced damage. These findings suggest that genetic fitness landscapes are dynamic and contain valuable time and condition-specific information. An additional tool built by our lab, systematic-gene-to-phenotype arrays (SGPA), permits interrogation of genetic landscapes behind given phenotype. This proposal uses GEoDE and SGPA to build a complete model of the dynamic response to DNA damage, first via genome-wide screens in the genetically tractable model organism S. cerevisiae (Aim 1), and then via targeted screens in human cell lines (Aim 2). The ultimate goal is to identify specific therapeutic targets and treatment windows in the DNA damage response pathway.